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在韩国红鱼(Sebastes schlegeli)性腺发育过程中β型雌激素受体(ERβ1 和 ERβ2)的分子克隆与表达分析。

Molecular cloning and expression analysis of estrogen receptor betas (ERβ1 and ERβ2) during gonad development in the Korean rockfish, Sebastes schlegeli.

机构信息

Fisheries College, Ocean University of China, Qingdao 266003, China.

出版信息

Gene. 2013 Jul 1;523(1):39-49. doi: 10.1016/j.gene.2013.03.109. Epub 2013 Apr 12.

Abstract

Estrogen receptors (ER) play a crucial role in mediation of estrogen activities. Here we report the isolation and expression analysis of ERβ1 and ERβ2 from ovary Korean rockfish (Sebastes schlegeli). were isolated using reverse transcription-polymerase chain reaction (PCR) and rapid amplification of cDNA ends procedures. The cDNA of this study, ERβ1 (588 amino acids) and ERβ2 (659 amino acids) were identified using reverse-transcriptase PCR (RT-PCR) and rapid amplification of cDNA ends procedures. Structural analysis showed both ERβs contain six typical nuclear receptor-characteristic domains. Phylogenetic analysis indicated that Korean rockfish ERβs were highly conserved among teleost. RT-PCR confirmed that the ERβs were widely distributed in both gonads and extra gonadal tissues. Further, we analyzed the expression patterns of male and female S. schlegeli during the reproductive cycle using quantitative real-time PCR (qRT-PCR). The results showed that the highest expression levels were observed in testis at immature sperm stage for both of KrERβ1 and KrERβ2. For female, the expressions of KrERβ1 and KrERβ2 were significantly higher in the ovary at the early-oocyte stage. Cloning these two ERβ subtypes in the Korean rockfish, together with the information on expression levels in adult fish has given us the foundation to investigate their possible role in brain-pituitary-gonad neuroendocrine axis in future studies.

摘要

雌激素受体 (ER) 在介导雌激素活性方面发挥着至关重要的作用。在这里,我们报告了从卵巢朝鲜黑石斑鱼 (Sebastes schlegeli) 中分离和表达分析 ERβ1 和 ERβ2。使用反转录 - 聚合酶链反应 (PCR) 和快速扩增 cDNA 末端程序分离。本研究的 cDNA,ERβ1(588 个氨基酸)和 ERβ2(659 个氨基酸)通过反转录 -PCR(RT-PCR)和快速扩增 cDNA 末端程序鉴定。结构分析表明,两种 ERβ 都包含六个典型的核受体特征结构域。系统发育分析表明,朝鲜黑石斑鱼 ERβ 在硬骨鱼中高度保守。RT-PCR 证实 ERβ 在性腺和性腺外组织中广泛分布。此外,我们使用定量实时 PCR (qRT-PCR) 分析了雄性和雌性 S. schlegeli 在生殖周期中的表达模式。结果表明,在未成熟精子阶段的睾丸中,KrERβ1 和 KrERβ2 的表达水平最高。对于雌性,在早期卵母细胞阶段的卵巢中,KrERβ1 和 KrERβ2 的表达水平显著更高。在朝鲜黑石斑鱼中克隆这两种 ERβ 亚型,并结合成年鱼类的表达水平信息,为我们未来研究它们在脑垂体性腺神经内分泌轴中的可能作用奠定了基础。

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